酶解蚕豆蛋白制备降胆固醇肽及其响应面优化
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摘要
为了得到具有较高降胆固醇活性的蚕豆肽,对蚕豆蛋白的酶解条件进行了研究。通过比较6种蛋白酶对蚕豆蛋白的酶解程度以及降胆固醇能力,选择碱性蛋白酶进一步优化酶解条件。以水解度(DH)与降胆固醇能力为指标,利用单因素试验和响应面试验优化确定了酶解蚕豆蛋白制备降胆固醇肽的工艺参数。结果表明,最佳酶解参数为:酶解温度40℃,p H 9.0,酶解时间60 min,加酶量8 000 U/g,料液比1∶20。验证试验后得到的水解度为14.29%蚕豆多肽产品,其对胆酸钠、甘氨胆酸钠、牛磺胆酸钠抑制率分别为52.98%、31.87%、25.17%,与理论模型预测值52.49%、31.44%、24.76%基本相符。
The hypocholesterolemic peptides from broad bean protein by enzymatic hydrolysis were investigated.In order to obtain broad bean peptides with high cholesterol-lowering activity,six proteinases were compared on the degree of hydrolysis( DH) of broad bean protein and the hypocholesterolemic abilitity,and alkaline protease was employed.Then the DH and hypocholesterolemic ability were used as indicators,single factor experiment and response surface methodology were applied to get the optimal enzymatic hydrolysis parameters of broad bean protein.The results showed that the optimal enzymatic hydrolysis parameters for alkaline protease were obtained as follows: enzymolysis temperature 40 ℃,pH 9.0,enzymolysis time 60 min,dosage of enzyme 8 000 U/g and ratio of material to liquid 1∶ 20.Under the optimal conditions,broad bean peptides with 14.29% DH could be obtained,and its inhibition rates on sodium cholate,sodium glycine and sodium taurocholate were predicted 52.49% ,31.44% and 24.76% ,and the validation tests gave 52.98% ,31.87% and 25.17% ,respectively.
The hypocholesterolemic peptides from broad bean protein by enzymatic hydrolysis were investigated.In order to obtain broad bean peptides with high cholesterol-lowering activity,six proteinases were compared on the degree of hydrolysis( DH) of broad bean protein and the hypocholesterolemic abilitity,and alkaline protease was employed.Then the DH and hypocholesterolemic ability were used as indicators,single factor experiment and response surface methodology were applied to get the optimal enzymatic hydrolysis parameters of broad bean protein.The results showed that the optimal enzymatic hydrolysis parameters for alkaline protease were obtained as follows: enzymolysis temperature 40 ℃,pH 9.0,enzymolysis time 60 min,dosage of enzyme 8 000 U/g and ratio of material to liquid 1∶ 20.Under the optimal conditions,broad bean peptides with 14.29% DH could be obtained,and its inhibition rates on sodium cholate,sodium glycine and sodium taurocholate were predicted 52.49% ,31.44% and 24.76% ,and the validation tests gave 52.98% ,31.87% and 25.17% ,respectively.
引文
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[22]苏海玲,张海英,韩涛,等.蚕豆蛋白酶解工艺及响应面法的优化[J].中国食品学报,2013,13(5):39-46.
[23]李慧,丁轲,韩涛,等.响应面法优化蚕豆蛋白酶解工艺条件的研究[J].中国食物与营养,2015,21(4):31-35.
[24]宋晓敏.蚕豆蛋白的提取、功能特性及酶水解研究[D].天津:天津商业大学,2013.
[25]吴继美.粮食水溶性蛋白结合胆酸盐能力及胆酸盐免疫检测方法的研究[D].广州:暨南大学,2014.
[26]周小理,黄琳,周一鸣.苦荞水溶性蛋白体外吸附胆酸盐能力的研究[J].食品科学,2011,32(23):77-81.
[2]宋晓敏,李素芬,刘建福,等. 20种蚕豆样品蛋白质含量与其蛋白组分的分子质量[J].中国粮油学报,2013,28(12):42-46.
[3]刘凤芹.冀西北蚕豆优异资源筛选评价[D].北京:中国农业科学院,2009.
[4]焦凌梅,袁唯.蚕豆中抗营养因子的研究[J].粮油加工与食品机械,2004(2):51-53.
[5]CABANOS C,EKYO A,AMARI Y,et al. High-level production of lactostatin,a hypocholesterolemic peptide,in transgenic rice using soybean A1aB1b as carrier[J].Transg Res,2013,22(3):621-629.
[6]GONZLEZ-ORTEGA L O,LPEZ-LIMON A R,MORALES-DOMINGUEZ J F,et al. Production and purification of recombinant hypocholesterolemic peptides[J].Biotech Lett,2015,37(1):41-54.
[7]PAK V V,KOO M,LEE N,et al. Structure-activity relationships of the peptide Ile-Ala-Val-Pro and its derivatives revealed using the semi-empirical AM1 method[J]. Chem Nat Compd,2005,41(4):454-460.
[8]高学飞.乳清蛋白源降胆固醇活性肽的制备及其生物学功能的研究[D].合肥:安徽农业大学,2006.
[9]王茵,苏永昌,吴靖娜,等.紫菜多肽降血脂及抗氧化作用的研究[J].食品工业科技,2013,34(16):334-337.
[10]宋玲钰.花生降胆固醇肽的制备及饮料开发[D].山东泰安:山东农业大学,2016.
[11]YAO Y,ZHANG J,HUI H,et al. Preparation and bioactivity of chickpea hypocholesterolemic peptides[J]. J Chin Cereal Oils Assoc,2015,30(1):33-38.
[12]于娜.卵黄多肽的分离纯化及降血脂活性研究[D].沈阳:沈阳农业大学,2012.
[13]文浪.具有胆酸盐吸附作用的苦荞寡肽的制备及活性评价[D].上海:上海应用技术大学,2016.
[14]黄昆.山杏仁多肽的制备及其对胆酸盐络合能力的研究[D].北京:北京林业大学,2013.
[15]白利峰,刘金双,周培根.扇贝边多肽提取物对高脂血症大鼠血脂以及脂质氧化的影响[J].安徽农业大学学报,2008,35(4):615-618.
[16]江锟.鲈鱼蛋白的酶解工艺优化及活性肽的功能特性研究[D].武汉:华中农业大学,2013.
[17]蓝建京,刘旭辉,伍善广,等.酶法制取丝素降胆固醇肽的工艺研究[J].安徽农业科学,2011,39(21):13119-13121.
[18]MEGAS C,PEDROCHE J,DEL M Y M,et al. Sunflower protein hydrolysates reduce cholesterol micellar solubility[J]. Plant Food Hum Nutr,2009,64(2):86-93.
[19]MARQUES M R,FONTANARI G G,PIMENTA D C,et al. Proteolytic hydrolysis of cowpea proteins is able to release peptides with hypocholesterolemic activity[J].Food Res Int,2015,77:43-48.
[20]郑睿.降胆固醇亚麻籽蛋白酶解肽的制备及结构表征[D].呼和浩特:内蒙古农业大学,2016.
[21]刘淳,张海英,韩涛,等. Alcalase碱性蛋白酶酶解蚕豆蛋白的研究[J].中国粮油学报,2011,26(12):29-33.
[22]苏海玲,张海英,韩涛,等.蚕豆蛋白酶解工艺及响应面法的优化[J].中国食品学报,2013,13(5):39-46.
[23]李慧,丁轲,韩涛,等.响应面法优化蚕豆蛋白酶解工艺条件的研究[J].中国食物与营养,2015,21(4):31-35.
[24]宋晓敏.蚕豆蛋白的提取、功能特性及酶水解研究[D].天津:天津商业大学,2013.
[25]吴继美.粮食水溶性蛋白结合胆酸盐能力及胆酸盐免疫检测方法的研究[D].广州:暨南大学,2014.
[26]周小理,黄琳,周一鸣.苦荞水溶性蛋白体外吸附胆酸盐能力的研究[J].食品科学,2011,32(23):77-81.